Dendritic cells (DC) generation from peripheral blood mononuclear cells obtained from jaundiced patients with pancreatic adenocarcinoma. / Análise de células mononucleares mantidas em cultura em meio propício para geração de células dendríticas obtidas de pacientes com câncer de pâncreas.

AUTOR(ES)
DATA DE PUBLICAÇÃO

2008

RESUMO

Pancreatic adenocarcinoma (PAdc) is an aggressive malignancy with poor prognosis, urging for improved or new therapeutic strategies. DC-based vaccination is one of such promising approaches. DC are the most potent antigen-presenting cells and central to the induction and maintenance of an immune response. However, in cancer patients DC generation and function may be deficient, imposing an obstacle to the success of their use. Here, we describe the in vitro generation of DC from peripheral blood mononuclear cells (PBMC) obtained from jaundiced patients with PAdc and, also, the effect of jaundiced plasma (JP) in the phenomenon. PBMC were separated from blood obtained from 10 patients and 10 healthy controls over a density gradient. Adherent cells were cultured with GM-CSF and IL-4 (50ng/mL) for 7 days. On the 5th day, TNF-a (50ng/mL) was added for DC activation. Cultures were performed in 10% JP or normal plasma (NP). Non-adherent cells were harvested at day 7, labeled with FITC- or PE-conjugated monoclonal antibodies against CD86, CD11c, CD14, HLA-DR and analyzed by flow cytometry. Patientscells, cultured in 10% JP, compared to healthy donorscells, cultured in 10% NP, had a significantly (p<0.05) lower expression of CD86 and HLA-DR. It is noteworthy that cells generated from patientsPBMC did not express CD11c, while from those derived from healthy donorscells did so. The presence of JP in healthy donorscells cultures caused a significant decrease in the percentage of HLA-DR+, CD11c+ and CD86+ cells. Finally, when patientsPBMC were cultured in NP, a significant increase in HLA-DR and in CD86 expression occurred. MLR assays also demonstrated that cells from jaundice patients had decreased capacity to stimulate alloestimulation of lymphocytes when compared to healthy donors. These data indicate a significant alteration in the patientsPBMC ability to differentiate into DC in vitro, a phenomenon that seems to depend both on soluble factors present in plasma and on the cells, themselves.

ASSUNTO(S)

cancer pancreas cell dendritica célula dendrítica jaundice icterícia câncer pâncreas

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